Dosio F, Brusa P, Crosasso P, Fruttero C, Cattel L, Bolognesi A
Laboratorio di Chimica Farmaceutica Applicata, Dipartimento di Scienza e Technologia del Farmaco, Torino, Italy.
Farmaco. 1996 Jul;51(7):477-82.
This study describes the synthesis of a panel of immunotoxins made by a non-covalent interaction between a monoclonal antibody derivatized with a dichlorotriazinic dye and six different ribosomal inhibitor proteins: colocin 1, momorcochin, momordin, bryodin, saporin 6 and PAP-S. The scheme of preparation showed several advantages respect to commercially available heterobifunctional cross-linkers, such as an higher overall yield of production and the homogeneity of the obtained conjugates. Nevertheless this procedure allowed the synthesis of immunoconjugates only for the four glycosilated RIPs since the not glycosilated ones saporin 6 and PAP-S precipitated in the presence of the dye. The non covalent linkage did not significantly affect the toxic activity of the glycosilated RIPs, as shown by their antitumoral activity on three different cell lines: HT-29 and A431 as target and MeWo as control. Furthermore, the monoclonal antibody cell-antigen recognition was preserved if a maximum derivatization ratio of 4 between the antibody and the dye was applied. The described original procedure may be of general application to prepare panels of immunotoxin for clinical use avoiding the expected RIP-related immune reaction.
本研究描述了一组免疫毒素的合成，该免疫毒素由用二氯三嗪染料衍生化的单克隆抗体与六种不同的核糖体抑制蛋白通过非共价相互作用制成：大肠菌素1、苦瓜素、巴豆苷、苔藓抑素、皂草素6和PAP-S。制备方案相对于市售的异双功能交联剂显示出几个优点，例如更高的总产率和所得缀合物的均一性。然而，该方法仅能合成四种糖基化核糖体失活蛋白的免疫缀合物，因为非糖基化的皂草素6和PAP-S在染料存在下会沉淀。非共价连接并未显著影响糖基化核糖体失活蛋白的毒性活性，这在它们对三种不同细胞系（作为靶标的HT-29和A431以及作为对照的MeWo）的抗肿瘤活性中得到了体现。此外，如果抗体与染料之间的最大衍生化率为4，则单克隆抗体的细胞-抗原识别得以保留。所描述的原始方法可能具有普遍适用性，可用于制备用于临床的免疫毒素组，避免预期的与核糖体失活蛋白相关的免疫反应。
